We propose to establish that tumor-specific antigens exist in human cancer. We will study a very prevalent form of cancer, squamous cell carcinoma (SCC). To characterize tumor antigens in human squamous carcinoma, we will: (1)\establish a comprehensive serum bank containing serial serum specimens from many patients; (2)\establish new cell lines of human squamous carcinoma and corresponding normal fibroblasts from the same patients; (3)\preserve in our cell bank all new cultures; (4)\characterize completely all new cell lines and make them available for study by others; (5)\survey the autologous serum-tumor cell combinations with sensitive mixed hemadsorption and immune adherence assays for antibody reactive with cell surface antigens; and (6)\establish antibody specificity and antigen distribution by adsorption analysis. Forty-one new SCC cell lines have been established from 36 patients. A characteristic membrane antigen phenotype of SCC lines distinguishes these lines from other tumor types as well as individual phenotypes that distinguish them from each other. SCC lines express: (1)\the epidermal cell-specific pemphigus antigen; (2)\blood group antigen corresponding to the donor blood type; and (3)\HLA heavy chain and beta2microglobulin. Reactivity with antibody to pemphigoid antigen is common with SCC lines but not with other cell types. Monoclonal antibodies raised to UM-SCC-1 have been used to characterize additional SCC antigens. These include the type II-H blood group antigens detected by antibody G10 and expressed by greater than 90% of SCC lines and 16/16 tumor biopsies studied by immunoperoxidase assays. An epithelial basal cell antigen detected by antibody A9 is displayed in an orientation-dependent fashion in cultured normal squamous cells but on all surfaces by malignant squamous cells in culture. Autologous antibody reactivity is detectable in serum from many cell line donors. Absorption analysis of serum from one patient with high antibody levels defines an antigen present on two autologous SCC tumor lines but not on normal fibroblasts from this patient. The antigen also appears to be present on some other SCC lines but not on melanoma lines. Studies underway are: (1)\continued characterization of autologous serum-defined antigens; (2)\characterization of factors mediating growth promotion of SCC cells in vitro; (3)\sensitivity tests of SCC cells to chemotherapeutic agents in vitro; (4)\assessment of which SCC lines grow in nude mice; and (5)\expression of steroid hormone receptors by laryngeal carcinomas and the in vitro response of hormone receptor positive cells to antihormones. (AG)